The application of hematoxylin must be carefully controlled to avoid over-staining, which can obscure the red lipid droplets. The Staining Procedure The standard Oil Red O staining procedure involves two primary solutions: a working stain solution and a rinsing solution.
Frozen Section Oil Red O Staining Guide
Fresh tissues should be snap-frozen in optimal cutting temperature compound and stored at low temperatures until sectioning. Sections are typically cut at a thickness of 5 to 10 micrometers.
After counterstaining, the sections are rinsed with water and mounted using an aqueous mounting medium. The working solution is prepared by dissolving Oil Red O powder in propylene glycol and filtering the mixture to ensure clarity.
Frozen Section Oil Red O Staining Guide
It is important to note that oil-based mounting media must be avoided as they can dissolve the Oil Red O dye, leading to fading and loss of the staining signal. Researchers can define specific regions of interest within the tissue, such as atherosclerotic plaques or hepatocytes, and calculate the percentage of area occupied by lipid droplets.
More About Oil red o staining
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